File No: LTD/1414
September 2009
NATIONAL INDUSTRIAL CHEMICALS NOTIFICATION AND ASSESSMENT SCHEME
(NICNAS)
FULL PUBLIC REPORT
Y-513
This Assessment has been compiled in accordance with the provisions of the Industrial Chemicals (Notification and
Assessment) Act 1989 (Cwlth) (the Act) and Regulations. This legislation is an Act of the Commonwealth of Australia.
The National Industrial Chemicals Notification and Assessment Scheme (NICNAS) is administered by the Department
of Health and Ageing, and conducts the risk assessment for public health and occupational health and safety. The
assessment of environmental risk is conducted by the Department of the Environment, Water, Heritage and the Arts.
For the purposes of subsection 78(1) of the Act, this Full Public Report may be inspected at our NICNAS office by
appointment only at 334-336 Illawarra Road, Marrickville NSW 2204.
This Full Public Report is also available for viewing and downloading from the NICNAS website or available on
request, free of charge, by contacting NICNAS. For requests and enquiries please contact the NICNAS Administration
Coordinator at:
Street Address:
Postal Address:
TEL:
FAX
Website:
Director
NICNAS
334 - 336 Illawarra Road MARRICKVILLE NSW 2204, AUSTRALIA.
GPO Box 58, SYDNEY NSW 2001, AUSTRALIA.
+ 61 2 8577 8800
+ 61 2 8577 8888
www.nicnas.gov.au
TABLE OF CONTENTS
FULL PUBLIC REPORT .............................................................................................................................................. 4
1.
APPLICANT AND NOTIFICATION DETAILS ............................................................................... 4
2.
IDENTITY OF CHEMICAL ............................................................................................................... 4
3.
COMPOSITION .................................................................................................................................. 4
4.
PHYSICAL AND CHEMICAL PROPERTIES .................................................................................. 4
5.
INTRODUCTION AND USE INFORMATION ................................................................................ 5
6.
HUMAN HEALTH IMPLICATIONS ................................................................................................ 6
6.1
Exposure assessment .................................................................................................................. 6
6.1.1
Occupational exposure.................................................................................................................... 6
6.1.2. Public exposure. .............................................................................................................................. 7
6.2.
Human health effects assessment................................................................................................ 7
6.3.
Human health risk characterisation .......................................................................................... 10
6.3.1. Occupational health and safety ..................................................................................................... 10
6.3.2. Public health ................................................................................................................................. 11
7.
ENVIRONMENTAL IMPLICATIONS ............................................................................................ 11
7.1
Environmental Exposure & Fate Assessment ........................................................................... 11
7.1.1
Environmental Exposure ............................................................................................................... 11
7.1.2
Environmental fate. ....................................................................................................................... 11
7.1.3
Predicted Environmental Concentration (PEC) ............................................................................ 12
7.2
Environmental effects assessment ............................................................................................ 12
7.2.1
Predicted No-Effect Concentration ............................................................................................... 12
7.3
Environmental risk assessment ................................................................................................. 12
8.
CONCLUSIONS AND REGULATORY OBLIGATIONS .............................................................. 12
APPENDIX A: PHYSICAL AND CHEMICAL PROPERTIES ........................................................................................... 15
APPENDIX B: TOXICOLOGICAL INVESTIGATIONS ................................................................................................... 18
B.1.
Acute toxicity – oral ................................................................................................................. 18
B.2.
Acute toxicity – dermal............................................................................................................. 18
B.3.
Irritation – skin ......................................................................................................................... 19
B.4.
Irritation – eye .......................................................................................................................... 19
B.5.
Skin sensitisation – mouse local lymph node assay (LLNA) ................................................... 20
B.6.
Repeat dose toxicity.................................................................................................................. 20
B.7.
Genotoxicity – bacteria ............................................................................................................. 21
B.8.
Genotoxicity – in vitro.............................................................................................................. 22
APPENDIX C: ENVIRONMENTAL FATE AND ECOTOXICOLOGICAL INVESTIGATIONS ................................................ 24
C.1.
Environmental Fate ................................................................................................................... 24
C.1.1.
Ready biodegradability ................................................................................................................ 24
C.1.2.
Bioaccumulation .......................................................................................................................... 24
C.2.
Ecotoxicological Investigations. ............................................................................................... 24
C.2.1.
Acute toxicity to fish.................................................................................................................... 24
C.2.2.
Acute toxicity to aquatic invertebrates......................................................................................... 25
C.2.3.
Algal growth inhibition test ......................................................................................................... 25
C.2.4.
Inhibition of microbial activity .................................................................................................... 26
BIBLIOGRAPHY ...................................................................................................................................................... 27
Created on 9/22/2009 6:01:00 PM
PM
Last Saved 10/26/2009 1:10:00
September 2009
NICNAS
FULL PUBLIC REPORT
Y-513
1.
APPLICANT AND NOTIFICATION DETAILS
APPLICANT(S)
DIC Australia Pty Ltd (ABN 12 000 079 550)
42 Sunmore Close, Heatherton VIC 3202
NOTIFICATION CATEGORY
Limited-small volume: Chemical other than polymer (1 tonne or less per year).
EXEMPT INFORMATION (SECTION 75 OF THE ACT)
Data items and details claimed exempt from publication:
Chemical name, Other names, CAS number, Molecular formula, Structural formula, Molecular weight,
Spectral data, Method of detection and determination, Purity, Additives, Import volume, Identity of recipients
VARIATION OF DATA REQUIREMENTS (SECTION 24 OF THE ACT)
No variation to the schedule of data requirements is claimed.
PREVIOUS NOTIFICATION IN AUSTRALIA BY APPLICANT(S)
None
NOTIFICATION IN OTHER COUNTRIES
None
2.
IDENTITY OF CHEMICAL
MARKETING NAME(S)
Y-513
Symuler Fast Yellow 4407NF (containing 3-8% notified chemical)
MOLECULAR WEIGHT
>500 Da
ANALYTICAL DATA
Reference NMR, IR, HPLC, MS(ESI), ICP-AES, UV spectra were provided.
3.
COMPOSITION
The notified chemical is a reaction mixture consisting of several structurally related azo components derived
from 3,3’-dichlorobenzidine.
DEGREE OF PURITY
>95%
HAZARDOUS IMPURITIES/RESIDUAL MONOMERS
None. The notifier has indicated that they expect that the notified chemical does not contain impurities of 3,3’dichlorobenzidine at levels greater than 10ppm.
4.
PHYSICAL AND CHEMICAL PROPERTIES
APPEARANCE AT 20ºC AND 101.3 kPa: Yellow powder
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Property
Melting Point/Freezing Point
Boiling Point
Density
Vapour Pressure
Water Solubility
Hydrolysis as a Function of pH
Partition Coefficient
(n-octanol/water)
Adsorption/Desorption
Dissociation Constant
Particle Size
Flash Point
Value
Decomposes at approx. 300°C
Decomposes at approx. 300°C
1450 kg/m3 at 20oC
< 1.33 x 10-11 kPa at 20oC
< 0.36 mg/L at 20oC
Not measured, because of very low
water solubility.
log Pow > 6.5 at 20oC
Data Source/Justification
Measured
Measured
Measured
Measured
Measured
Expected to be stable to hydrolysis.
log Koc > 5.63 at 35C
Not measured, because of very low
water solubility.
Inhalable fraction (<100 m):
90.69 %
Respirable fraction (<10 m):
24.86 %
MMAD = 30.615 m
Not determined
Measured
Not expected to dissociate in the
environment.
Measured
Flammability
Not flammable
Autoignition Temperature
Not self-ignitable
Explosive Properties
Not explosive
Oxidising Properties
Not expected to be oxidising
* MMAD = Mass Median Aerodynamic Diameter
Measured
Test not conducted as
notified chemical is a solid
Measured
Measured
Measured
Estimated
DISCUSSION OF PROPERTIES
For full details of tests on physical and chemical properties, please refer to Appendix A.
Reactivity
The notified chemical is expected to be stable under normal conditions of use. The notified chemical does not
contain any structural indications of oxidising properties or other particular reactivity.
5.
INTRODUCTION AND USE INFORMATION
MODE OF INTRODUCTION OF NOTIFIED CHEMICAL (100%) OVER NEXT 5 YEARS
The notified chemical will be imported in yellow pigment at concentrations of 3-8% and in printing inks at
concentrations of 0.5%.
MAXIMUM INTRODUCTION VOLUME OF NOTIFIED CHEMICAL (100%) OVER NEXT 5 YEARS
Year
Tonnes
1
<1
2
<1
3
<1
4
<1
5
<1
PORT OF ENTRY
Sydney, Melbourne
TRANSPORTATION AND PACKAGING
The imported notified chemical will be transported by truck in Kraft paper bags with a laminated liner (10kg)
and in flexible containers (300kg).
Printing inks will be transported in cans (18L) and drums (200L), by truck.
USE
The notified chemical will be used as a pigment in printing inks. The products that it will be used to print paper
or film substrates including magazines, catalogues, food packaging (not direct food contact), etc.
OPERATION DESCRIPTION
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Formulation
The notified chemical (3-8% concentration) will be weighed either by counting the number of paper bags
(known to contain 10kg per bag) or by manual weighing under local ventilation for quantities smaller than
10kg. It will then be transferred by manual emptying of the paper bags into a hopper that is fitted with a dust
extractor (local ventilation equipment) together with a few other ingredients. Alternatively, for larger volumes
of pigment (3-8% notified chemical) that are imported in flexible 300kg containers, the contents will be
transferred into the hopper using automated procedures involving little direct handling by workers. From the
hopper the material will then be pumped into an enclosed mixing tank. It will be mixed in a closed tank with
other ingredients. At the completion of the mixing, the concentration of the notified chemical will be 0.5%. It
will then be transferred to a kneader or beads mill for processing in a closed system. This transfer will be
performed using a pump and hosing. During milling, quality control sampling will involve removing samples
using a sampling valve. After milling, the processed ink containing the notified chemical (0.5%) will be
transferred to packaging containers through a filter using a pump and hosing. Filters will be changed after this
transfer and washed with organic solvents. Equipment will be cleaned by circulating organic solvent.
End use
Printing using inks containing the notified chemical (0.5%) will be performed mainly using gravure printing
and also some using conventional offset printing processes (that may also include heat-set drying). Generally,
the ink containing the notified chemical (0.5%) will be transferred to the printing machine with a pump and
through a filter (if necessary). Filters will be removed and washed with organic solvents. The printing process
itself will be automated. For heat-set inks, the printed material will be briefly placed in a heatset oven that will
typically be at temperatures of 120oC (range of 100 - 130 oC). Any ink remaining on the printing machine will
be wiped off with a waste cloth soaked in organic solvents.
6.
HUMAN HEALTH IMPLICATIONS
6.1
6.1.1
Exposure assessment
Occupational exposure
NUMBER AND CATEGORY OF WORKERS
Category of Worker
Printing ink formulators
Mixing
Milling and packaging
Printing ink users
Charging
Number
Exposure Duration
(hours/day)
Exposure Frequency
(days/year)
2
2
30 minutes/day
2 hours/day
80 times/year
80 times/year
8
10 minutes/day
80 times/year
EXPOSURE DETAILS
Formulation
Inhalation exposure of workers to the notified chemical in powder form at concentrations of 3-8% may occur
when weighing and transferring pigments to the mixing tank during ink formulation processes. The use of dust
extractors and local ventilation systems is expected to collect most pigments containing the notified chemical
that may be scattered during these processes. EASE modelling predicts dust exposure of 0.16-0.4 mg/m3 (for
powders consisting of 8% of the notified chemical), assuming that effective local exhaust ventilation is
utilised. For a 70 kg worker, assuming an inhalation rate of 1.3 m3/h, 4 hour exposure and that the notified
chemical was all either absorbed or ingested via the mucociliary clearance mechanism, systemic exposure after
inhalation is estimated to be 0.01-0.03 mg/kg bw/day. This estimate assumes that no respiratory protection is
worn. However, the notifier has indicated that dust protective masks are expected to be worn during such
procedures.
The process of weighing and transferring pigments to the mixing tank may also result in dermal and ocular
exposure of workers to the notified chemical (3-8%). EASE modelling predicts ‘very low’ dermal exposure
during such processes. Workers are also expected to wear personal protective equipment such as gloves, safety
glasses, overalls, etc, that should further lower the potential for exposure.
Dermal and accidental ocular exposure of workers to the notified chemical at concentrations of 0.5% may
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occur during the occasional connecting and disconnecting of hoses from the mixing tank to the beads mill or
from the beads mill to packaging. EASE modelling of the pipe disconnection, cleaning and quality control
operations was performed to estimate dermal exposure of workers to the notified chemical. The following
assumptions were used for these estimates: direct handling, non-dispersive use (only used by workers with
knowledge of the processes and use of controls), and incidental contact level (assumed to be one event per
day). The predicted dermal exposure to the notified chemical is up to 0.0005 mg/cm2/day. This is equivalent to
up to 0.0003 mg/kg bw/day, based on dermal absorption of 10%, a surface area of 420 cm2 (equivalent to the
area of one hand or two half hands), and 70kg worker, and that the notified chemical is present at 0.5%
concentration (EC, 2003). Such exposure is expected to be lowered by the use of personal protective
equipment such as gloves, safety glasses and perhaps also protective masks.
Dermal and accidental ocular exposure of workers to the notified chemical at concentrations of 0.5% may also
occur during quality control sampling, cleaning of removed filters and other equipment. Such exposure is
expected to be lowered by the use of personal protective equipment such as gloves, safety glasses and perhaps
also protective masks.
The total systemic exposure after both inhalation and dermal exposure is estimated to be 0.0103 0.0303 mg/kg bw/day. The lower end of this range is considered to be a more appropriate estimate of
exposure when local exhaust ventilation is present and dermal protection is worn.
End use
Dermal and ocular exposure of workers to the notified chemical at concentrations of 0.5% may occur during
transfer of inks to the printing machine, cleaning of filters and printing equipment, and contact with printed
materials. Such exposure is expected to be lowered by the use of personal protective equipment such as gloves,
safety glasses and, where necessary, protective masks. Also, during contact with the printed material, the ink
will often be dry and thus the notified chemical should be trapped within a dried matrix and be unavailable for
exposure. The notifier has also stated that for some end use applications the notified chemical will be covered
by resin following its printing onto the substrate, which would minimise exposure.
6.1.2. Public exposure
The general public may make dermal contact with articles printed with the ink containing the notified
chemical. However, once the ink has dried, the notified chemical will be trapped within the dried matrix and is
not expected to be bioavailable. Whilst the potential for blooming and bleeding of the notified chemical from
the substrate may exist, it is expected that the inks containing the notified chemical will be appropriately
formulated with other ingredients and processed such as to minimise the tendency to bleed.
6.2.
Human health effects assessment
The results from toxicological investigations conducted on the notified chemical are summarised in the table
below. Details of these studies can be found in Appendix B.
Endpoint
Rat, acute oral toxicity
Rat, acute dermal toxicity
Rat, acute inhalation toxicity
Rabbit, skin irritation
Rabbit, eye irritation
Mouse, skin sensitisation – Local lymph node assay
Rat, repeat dose oral toxicity – 28 days.
Mutagenicity – bacterial reverse mutation
Genotoxicity – in vitro mammalian chromosome
aberration test
Result and Assessment Conclusion
low oral toxicity LD50 >2000 mg/kg bw
low dermal toxicity LD50 >2000 mg/kg bw
Not determined
non-irritating
slightly irritating
no evidence of sensitisation
NOAEL 1000 mg/kg bw/day
non mutagenic
non genotoxic
Note: there is a significant amount of toxicological information available on a chemical that is structurally
related to the two major components of the notified chemical (an azo pigment derived from 3,3’dichlorobenzidine). Where relevant, such data will be detailed in the below discussion, together with that on the
notified chemical itself.
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Toxicokinetics
The notified chemical has relatively high molecular weight (>500 Da), low water solubility (<0.36 mg/L) and
high partition coefficient (log P >6.5).
Based on its chemical properties (relatively high molecular weight, low water solubility and high partition
coefficient), absorption of the notified chemical through the skin is not expected to be significant (European
Commission 2003, NOTOX 2009b).
Systemic exposure following oral administration of the notified chemical is not expected to be significant as its
absorption from the gastrointestinal tract is likely to be limited by its water solubility and molecular weight. Any
uptake is likely to occur via micellular solubilisation, given its highly lipophilic nature and low water solubility.
Yellow stool in the acute and repeated dose oral studies suggests that the main route of excretion is through the
faeces, and therefore indicate that the majority of the notified chemical is not absorbed from the gastrointestinal
tract.
Studies on a chemical that is structurally related to the notified chemical have shown that it was not absorbed
following oral ingestion or dermal administration in rats, with the majority of the dose being excreted in the
faeces following oral administration, or remaining at the application site following dermal application (Exempt
reference 2,6).
The vast majority of the notified chemical (~91%) is of inhalable (< 100 µm) particle size and could be inhaled
into the upper respiratory tract. A significant portion (~25%) is also of small enough particle size (<10 µm) to
reach the lower respiratory tract (tracheobronchial and pulmonary regions). Larger particles of inhalable size are
expected to deposit in the nasopharyngeal region and be cleared by coughing/sneezing or be swallowed. Due to
the low water solubility of the notified chemical smaller particles lodging in the tracheobronchial region are
likely to be cleared by the mucociliary mechanism and swallowed. However, inhaled respirable particulates of
the notified chemical lodging in the pulmonary region may not be readily cleared due to its low water solubility.
There may be some potential for absorption across the respiratory tract epithelium due to its lipophilic nature. In
summary, higher concentrations of exposure may be expected to result in increased impairment of clearance
mechanisms (European Commission 2003, NOTOX 2009b).
Azo compounds may also break down to their component amines. The azo linkage is the most labile portion of
an azo colourant molecule, and it is readily enzymatically metabolised in mammals, including man (SCCNFP,
2002). Liver azo reductase enzymes reductively cleave the molecules into component amines. Some metabolism
of azo colourants may also occur in the cells of the bladder wall, and during percutaneous absorption. Intestinal
bacteria are also capable of catalysing reductive cleavage of the azo bond. Any cleavage of the notified chemical
that may occur in the gastrointestinal tract, though limited, will result in molecules with different physical and
chemical properties than the notified chemical and are expected to be absorbed to a greater extent. The
metabolites of the notified chemical are expected to be excreted via the bile or the urine (NOTOX 2009b).
Several studies on structurally related chemicals have suggested that the degree of breakdown is low. Following
oral administration to test animals these studies did not detect significant amounts of 3,3’-dichlorobenzidine in
the urine of the animals (Exempt references 1,3). A rat inhalation study revealed no detectable levels of 3,3’dichlorobenzidine in the urine or blood, suggesting that the test substance was not metabolically cleaved
following inhalation (Exempt reference 3). Levels of 3,3’-dichlorobenzidine and monoacetyl-3,3’dichlorobenzidine were below the detection limit in all urine samples obtained from textile workers who had
been exposed to dichlorobenzidine based pigments (Exempt reference 3).
Acute toxicity
The notified chemical was found to be of low acute oral and dermal toxicity in studies conducted on rats
(LD50 > 2,000 mg/kg bw).
The acute inhalation toxicity of the notified chemical has not been determined. Based on the acute inhalation
toxicity of a structurally related chemical, the notified chemical may be at least harmful via inhalation.
Irritation and Sensitisation.
The notified chemical was found to be non-irritating to the skin of rabbits. It was found to be slightly irritating to
rabbit eyes, with irritation that was not sufficient to warrant hazard classification.
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The notified chemical was not a skin sensitiser when tested up to a concentration of 25% in a mouse local lymph
node assay (maximum concentration that could technically be applied). Therefore, the notified chemical is
unlikely to cause skin sensitisation in exposed humans. This is further supported by human patch testing that
was performed with the azo pigment that is structurally related to the notified chemical (Exempt reference 10).
However, there are some reports of skin sensitisation of this structurally related chemical in humans (Exempt
reference 1).
Repeat Dose Toxicity.
The NOAEL in a 28 day oral repeat dose study in rats was determined to be 1000 mg/kg bw/day (the highest
tested dose). There were no toxicologically significant effects observed during the study.
Mutagenicity and Carcinogenicity.
Azo colourants are a concern for their potential induction of mutagenicity and carcinogenicity mainly through
the aromatic amines that are present as impurities in the colourants, or that arise from their azo reduction in or
outside of the body.
The notified chemical may be reductively cleaved to release one of the restricted aromatic amines specified in
the Appendix to EC Directive 76/769/EEC (EC, 2004), that is, 3,3’-dichlorobenzidine, CAS number 91-94-1,
which is a category 2 carcinogen.
The notified chemical did not appear to contain impurities of component arylamines based on the HPLC
information provided. The notifier has also indicated that they expect that the notified chemical does not contain
impurities of 3,3’-dichlorobenzidine at levels greater than 10ppm. As such, impurity levels are unlikely to
contribute to carcinogenicity of the notified chemical.
The extent of azo reduction through metabolism is not known, however it is thought that this is likely to be
lesser in pigments than in dyes due to reduced bioavailablity. It is noted that the EU and IARC lists benzidinebased azo dyes as being category 2 carcinogens (IARC 1987). However, this is not the case for 3,3’dichlorobenzidine based azo pigments (Exempt reference 3).
However, the German Commission for the Investigation of Health Hazards of Chemical Compounds in the
Work Area (DFG, 1988) states that “all azo colourants whose metabolism can liberate a carcinogenic aryl
amine are suspected of having carcinogenic potential.” It is also mentioned that if there are indications that the
colourant or its carcinogenic break down products are not bioavailable, the absence of risk needs to be proven
either experimentally, substantiated by biomonitoring, or suitable animal experiments performed to rule out
carcinogenic potential.
Amines may also be released during storage or processing. The USEPA considers dichlorobenzidine-based
pigments to be of concern for their potential release of 3,3’-dichlorobenzidine when used at temperatures in
excess of 200oC (when release of 3,3’-dichlorobenzidine may occur) (USEPA 2002).
The notified chemical was not mutagenic in bacteria, nor did it induce chromosomal aberrations in mammalian
cells in vitro. In both studies, precipitation of the test substance was observed at several of the concentrations
tested. An in vivo genotoxicity study was not provided.
The Ames test provided was performed in accordance with the modified procedure suggested for azo
compounds (Prival and Mitchell, 1982). The study utilised a reductive pre-incubation step (during which it is
expected that the azo colourant may be reduced to amine species) before the test is carried out to potentially
yield a greater detection of mutagenic azo species.
A structurally related chemical has been extensively tested for its genotoxicity. It was negative in several Ames
tests (including some studies that involved prior reduction of the azo pigment) (Exempt references 4,9) and at
least one chromosome aberration test (Exempt reference 8). However, one sister chromatid exchange test gave
equivocal results (Exempt reference 8) and it was genotoxic in hepatocytes in a comet assay (Exempt reference
7). One study tested metabolites of the azo colourant and found them to be strongly mutagenic (Exempt
reference 5). The lack of mutagenicity generally observed for the structurally related chemical has sometimes
been attributed to its lack of solubility in the test medium (Exempt references 1,3,9).
Several in vivo tests have also been performed on the carcinogenicity of a structurally related chemical. All such
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studies gave negative results (Exempt reference 4).
In summary, although the notified chemical may potentially be metabolised to a known carcinogen in vivo,
studies on the notified chemical itself, as well as a structurally related chemical, indicate that it is not likely to be
significantly bioavailable. In addition, in vivo carcinogenicity studies on this structurally related chemical
indicated that it was not an in vivo carcinogen. Overall, the data suggests that the notified chemical is not
expected to be mutagenic or carcinogenic, however as very low exposures can cause these effects, the potential
cannot be ruled out.
Health hazard classification
Based on the available data the notified chemical cannot be classified as hazardous under the Approved Criteria
for Classifying Hazardous Substances (NOHSC, 2004).
6.3.
Human health risk characterisation
6.3.1. Occupational health and safety
Respiratory effects
The acute and chronic effects after inhalation exposure to the notified chemical were not investigated, and
therefore an inhalation NOAEL can not be determined. Based on respiratory effects seen in the acute
inhalation studies of structurally related azo pigments of similar particle size distribution, the notified chemical
may pose both an acute and a chronic respiratory hazard. Repeated inhalation of airborne dusts of the notified
chemical, or inhalation of high airborne concentrations, may present some risk of lung overloading effects.
Inhalation exposure of workers to the notified chemical (concentrations up to 8%) may occur when handling
the notified chemical in solid form during formulation of the inks. Some of this airborne powder is expected to
be of respirable size. Since an inhalation NOAEL could not be determined the safety of the EASE estimated
atmospheric concentration after repeated exposure could not be established. Based on the potential for
respiratory effects after acute or chronic exposure the inhalation risk to workers handling the powder may be
significant.
The notifier has not specified the types of masks that will be worn by workers when formulating the notified
chemical into inks. If particle filter masks capable of filtering out particles of respirable size are worn by
formulation workers and cleaners and are used and fitted correctly, the exposure to the airborne notified
chemical, and therefore the risk to formulation and cleaning workers, will be significantly reduced.
Systemic effects
Dermal and inhalation exposure are the main routes by which workers may be exposed to the notified chemical
(at concentrations up to 8%) during formulation processes. EASE modelling predicted ‘very low’ levels of
dermal exposure assuming that effective local exhaust ventilation was in place.
Dermal exposure to the notified chemical at concentrations of 0.5% during pipe connection/disconnection,
cleaning and quality control operations may occur and some of the notified chemical may be absorbed into the
skin. It is noted that the use of PPE will act to further reduce exposure.
As the NOAEL value in a 28 day oral repeat dose toxicity study was determined to be greater than or equal to
the highest tested dose (1000 mg/kg bw/day), significant systemic effects are not expected to occur as a result
of dermal/inhalation exposure to the notified chemical.
In summary, the risk of systemic effects associated with dermal/inhalation exposure to the notified chemical
during formulation and end use is not considered to be unacceptable.
Mutagenicity/Carcinogenicity
Some potential for mutagenicity/carcinogenicity related to the carcinogenic metabolite, 3,3’-dichlorobenzidine,
cannot be ruled out.
Whilst breakdown of the pigment through metabolism in the body is possible it is believed that for insoluble
azo pigments such as the notified chemical this would be limited by the low solubility and thus low
bioavailability of the notified chemical (Exempt reference 3). This is also supported by substantial testing on a
structurally related chemical, including a negative carcinogenicity test.
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No component amines were identified as impurities in the notified chemical and the level of 3,3’dichlorobenzidine is expected to be low.
The engineering controls in place and personal protective equipment expected to be used by workers during
formulation and printing are expected to reduce exposure to a low level and consequently further minimise the
risk of mutagenicity or carcinogenicity.
The use of heat may result in degradation of the notified chemical to 3,3’-dichlorobenzidine (a category 2
carcinogen), particularly at temperatures in excess of 200oC (USEPA 2002). It is expected that the
temperatures of heatset ovens used for application of heat-set inks will not exceed 200oC and thus the notified
chemical is unlikely to degrade to 3,3’-dichlorobenzidine during such processes. Thus the risk associated with
end use of heat-set inks containing the notified chemical is unlikely to be significant.
Worker exposure to dried inks is likely to result in minimal exposure to the notified chemical, so the risk
presented by the notified chemical in this scenario is not expected to be significant.
In summary, the risk of mutagenic/carcinogenic effects associated with worker exposure to the notified
chemical is not considered to be unacceptable based on the use scenarios and controls in place to reduce
exposure.
6.3.2. Public health
Potential for mutagenic or carcinogenic effects related to the carcinogenic metabolite of the notified chemical,
3,3΄-dichlorobenzidine, cannot be ruled out.
The public will generally only have dermal exposure to dried inks containing the notified chemical, from
which it is not expected to be significantly bioavailable. Therefore the risk to the public is not considered to be
unacceptable.
7.
ENVIRONMENTAL IMPLICATIONS
7.1.
Environmental Exposure & Fate Assessment
7.1.1
Environmental Exposure
RELEASE OF CHEMICAL AT SITE
Minor releases from formulation (estimated as < 4 kg/year) are possible from spillages, container residues and
equipment cleaning.
RELEASE OF CHEMICAL FROM USE
Minor releases during use (estimated as < 6 kg/year) are possible from container residues and equipment
cleaning. The main source of environmental exposure is expected to arise from paper recycling, during which
the pigment will become detached from the fibres. The detached pigment would be expected to associate with
sludge because of its very low water solubility, with very little potential for aquatic exposure.
RELEASE OF CHEMICAL FROM DISPOSAL
Wastes from formulation and use are expected to be disposed of to landfill or by thermal decomposition.
7.1.2
Environmental fate
The notified chemical is expected to slowly degrade in situ following landfill disposal, as it has very low water
solubility and is resistant to biodegradation. It is not expected to bioconcentrate in fish, based on its structure
and the known behaviour of analogue pigments. While no analogue test reports have been provided, the low
bioconcentration factors (< 10) reported by the notifier are considered reliable based on the chemical structures
of the notified chemical and the nominated analogue, and the properties of organic pigments. In particular, the
notified chemical is expected to have a low potential for bioaccumulation because of its very limited affinity for
the lipid phase of living organisms, as has been discussed in the screening assessment of Pigment Red 187
(Environment Canada and Health Canada, 2008). For the details of the environmental fate studies please refer to
Appendix C.
FULL PUBLIC REPORT: LTD/1414
Page 10 of 27
September 2009
7.1.3
NICNAS
Predicted Environmental Concentration (PEC)
It is neither necessary nor meaningful to determine the PEC as no aquatic exposure is expected when the
notified chemical is used as proposed.
7.2.
Environmental effects assessment
The results from ecotoxicological investigations conducted on the notified chemical are summarised in the
table below. Details of these studies can be found in Appendix C.
Endpoint
Fish Toxicity
Daphnia Toxicity
Algal Toxicity
Inhibition of Bacterial Respiration
Result
EC50 > 0.024 mg/L
EC50 > 0.031 mg/L
EC50 > 0.0043
mg/L
EC50 > 100 mg/L
Assessment Conclusion
Not toxic to limit of water solubility
Not toxic to limit of water solubility
Not toxic to limit of water solubility
Not harmful
Aquatic toxicity testing was conducted at a single nominal concentration of 100 mg/L, with actual exposure
concentrations limited by the very low water solubility as tabulated above. No harmful effects were observed
from exposure to concentrations up to the limit of water solubility.
7.2.1
Predicted No-Effect Concentration
It is not possible to determine the PNEC as the median effect concentrations cannot be determined. The
notified chemical is not harmful to aquatic life at concentrations up to the solubility limit.
7.3. Environmental risk assessment
The notified chemical is not considered to pose a risk to the environment, as no aquatic exposure is expected
when it is used as proposed, and testing has shown that it is not harmful to aquatic life at concentrations up to
the solubility limit.
8.
CONCLUSIONS AND REGULATORY OBLIGATIONS
Hazard classification
Based on the available data the notified chemical cannot be classified as hazardous under the Approved Criteria
for Classifying Hazardous Substances [NOHSC:1008(2004)].
Human health risk assessment
Based on the proposed use scenarios and occupational controls in place to reduce exposure, the notified
chemical is not considered to pose an unacceptable risk to the health of workers.
When used in the proposed manner, the notified chemical is not considered to pose an unacceptable risk to
public health.
Environmental risk assessment
On the basis of the very low water solubility and the reported use pattern, the notified chemical is not considered
to pose a risk to the environment.
Recommendations
REGULATORY CONTROLS
Health Surveillance
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Page 11 of 27
September 2009

NICNAS
As the notified chemical may be harmful by inhalation, employers should consider carrying out health
surveillance of workers.
CONTROL MEASURES
Occupational Health and Safety

Employers should implement the following engineering controls to minimise occupational exposure to
the notified chemical as introduced in the powdered pigment:
 Local exhaust ventilation wherever weighing and addition to mixers occurs

Employers should implement the following safe work practices to minimise occupational exposure
during handling of the notified chemical as introduced in the powdered pigment:
 Avoid the formation of airborne dusts
 Avoid skin contact
 Regularly clean up any spills of the powdered pigment
 Minimisation of the use of heat during handling/processing/use of the notified chemical

Employers should implement the following safe work practices to minimise occupational exposure to
the notified chemical in end use inks:
 Situations under which the notified chemical may be subject to temperatures in excess of 200oC
should be avoided, particularly when heat-setting inks containing the notified chemical.

Measures should be taken to minimise the levels of 3,3’-dichlorobenzidine in the imported products
containing the notified chemical.

Employers should ensure that the following personal protective equipment is used by workers to
minimise occupational exposure to the notified chemical as introduced in powdered pigment:
 Respiratory protection sufficient for respirable particulates during processes where exposure to
dust is likely
 Gloves
 Coveralls
 Safety glasses

Employers should ensure that the following personal protective equipment is used by workers to
minimise occupational exposure to the notified chemical in printing inks:
 Gloves
 Coveralls
 Safety glasses
Guidance in selection of personal protective equipment can be obtained from Australian,
Australian/New Zealand or other approved standards.

A copy of the MSDS should be easily accessible to employees.

If products and mixtures containing the notified chemical are classified as hazardous to health in
accordance with the Approved Criteria for Classifying Hazardous Substances [NOHSC:1008(2004)]
workplace practices and control procedures consistent with provisions of State and Territory hazardous
substances legislation must be in operation.
Disposal

The notified chemical should be disposed of to landfill.
Emergency procedures

Spills or accidental release of the notified chemical should be handled by containment, collection and
subsequent safe disposal.
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Page 12 of 27
September 2009
NICNAS
Regulatory Obligations
Secondary Notification
This risk assessment is based on the information available at the time of notification. The Director may call for
the reassessment of the chemical under secondary notification provisions based on changes in certain
circumstances. Under Section 64 of the Industrial Chemicals (Notification and Assessment) Act (1989) the
notifier, as well as any other importer or manufacturer of the notified chemical, have post-assessment regulatory
obligations to notify NICNAS when any of these circumstances change. These obligations apply even when the
notified chemical is listed on the Australian Inventory of Chemical Substances (AICS).
Therefore, the Director of NICNAS must be notified in writing within 28 days by the notifier, other importer or
manufacturer:
(1)
Under Section 64(1) of the Act; if
 the importation volume exceeds one tonne per annum notified chemical;
 further information becomes available as to the mutagenic/carcinogenic potential of the notified
chemical or its potential for breakdown;
 the notified chemical is proposed to be used in printing inks for use by consumers;
or
(2)
Under Section 64(2) of the Act; if
 the function or use of the chemical has changed from a pigment used in printing inks, or is likely to
change significantly;
 the amount of chemical being introduced has increased from 1 tonne per annum, or is likely to
increase, significantly;
 the chemical has begun to be manufactured in Australia;
 additional information has become available to the person as to an adverse effect of the chemical
on occupational health and safety, public health, or the environment.
The Director will then decide whether a reassessment (i.e. a secondary notification and assessment) is required.
Material Safety Data Sheet
The MSDS of the notified chemical (and products containing the notified chemical) provided by the notifier
were reviewed by NICNAS. The accuracy of the information on the MSDS remains the responsibility of the
applicant.
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Page 13 of 27
September 2009
NICNAS
APPENDIX A: PHYSICAL AND CHEMICAL PROPERTIES
Melting Point/Freezing Point
Method
Remarks
Test Facility
OECD TG 102 Melting Point/Melting Range.
EC Directive 92/69/EEC A.1 Melting/Freezing Temperature.
Differential scanning calorimetry (DSC).
Reaction and/or decomposition of the test substance were observed above approximately
300oC. Melting was not observed below the temperature at which reaction and/or
decomposition started.
NOTOX (2009a)
Boiling Point
Method
Remarks
Test Facility
Decomposes at approx. 300°C
OECD TG 103 Boiling Point.
EC Directive 92/69/EEC A.2 Boiling Temperature.
Differential scanning calorimeter (DSC).
Reaction and/or decomposition of the test substance were observed above approximately
300oC. Boiling was not observed below the temperature at which reaction and/or
decomposition started.
NOTOX (2009a)
1450 kg/m3 at 20oC (solid)
Density
Method
Remarks
Test Facility
OECD TG 109 Density of Liquids and Solids.
EC Directive 92/69/EEC A.3 Relative Density.
Gas comparison stereopycnometer.
Average of two experiments.
NOTOX (2009a)
< 1.33 X 10-11 kPa at 20oC.
Vapour Pressure
Method
Remarks
Test Facility
OECD TG 104 Vapour Pressure.
EC Directive 92/69/EEC A.4 Vapour Pressure.
Isothermal thermogravimetric effusion method.
The vapour pressure of the test substance was found to be below the vapour pressure of
benzo(ghi)perylene.
NOTOX (2009a)
< 3.6 x 10-4 g/L at 20oC
Water Solubility
Method
Remarks
Test Facility
OECD TG 105 Water Solubility.
EC Directive 92/69/EEC A.6 Water Solubility.
Although the column elution method is recommended for determination of water
solubility at such low concentrations, this was precluded by the low solubility of the
notified chemical in volatile solvents. The test relied on a non-specific analytical method
(TOC) because the low solubility in water and organic solvents precluded a specific
method such as HPLC. The measured solubility may therefore be an overestimate.
NOTOX (2009a)
Hydrolysis as a Function of pH
Remarks
Decomposes at approx. 300°C
Not measured.
Hydrolytic stability could not be measured because of the very low aqueous solubility
and the lack of a sufficiently sensitive and specific analytical method. The absence of
transformation in the biodegradation test indicates that the notified chemical can be
expected to be hydrolytically stable, consistent with its structure.
Partition Coefficient (noctanol/water)
FULL PUBLIC REPORT: LTD/1414
log Pow > 6.5 at 20oC
Page 14 of 27
September 2009
Method
Remarks
Test Facility
NICNAS
OECD TG 117 Partition Coefficient (n-octanol/water), HPLC method.
EC Directive 92/69/EEC A.8 Partition Coefficient.
HPLC Method, using acidified tetrahydrofuran as mobile phase. The notified chemical
eluted from the column after the reference substance DDT.
NOTOX (2009a)
Adsorption/Desorption
– screening test
Method
Remarks
Test Facility
OECD 121 Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage
Sludge using High Performance Liquid Chromatography
EC Directive 2001/59/EC C.19 Estimation of the Adsorption Coefficient (Koc) on Soil
and on Sewage Sludge using High Performance Liquid Chromatography
No test substance peak was observed during isocratic elution, but one test substance
related peak eluted at 14.3 minutes (twice the time for isocratic elution of DDT) under
gradient elution.
NOTOX (2009a)
Dissociation Constant
Remarks
log Koc > 5.63 at 35C
Not measured
Measurement was precluded by the very low aqueous solubility and the lack of a
sufficiently sensitive and specific analytical method. The notified chemical is not expected
to dissociate under environmental conditions, based on the structure.
Particle Size
Method
OECD TG 110 Particle Size Distribution/Fibre Length and Diameter Distributions.
Range (m)
< 2.785
< 10
< 25.424
< 95.275
Remarks
Test Facility
Samples of the test substance were dispersed in kerosene and five runs performed to
ensure repeatability of results.
Mass Median Aerodynamic Diameter (MMAD) = 30.615 m
Chilworth (2008)
Flammability
Method
Remarks
Test Facility
Not highly flammable
EC Directive 92/69/EEC A.10 Flammability (Solids).
The flame of a gas burner ignited the test substance pile. In contact with the flame, the test
substance glowed, burned with a yellow flame, produced grey smoke and turned into a
charred residue. After removal of the ignition source, the flame extinguished after 10
seconds and no more glowing or propagation of combustion (e.g. by smouldering) was
observed. Since the test substance did not burn with flame or smoulder over 200 mm
within 4 minutes was not considered to be highly flammable.
NOTOX (2009a)
Autoignition Temperature
Method
Remarks
Mass (%)
10
24.86
50
90
Not self-ignitable
92/69/EEC A.16 Relative Self-Ignition Temperature for Solids.
An exothermic reaction was observed to occur during the experiment, though the
temperature of the sample during this reaction did not reach 400oC (as required by the test
guideline). After the experiment, a partially charred, white residue was observed in the
cube. No test substance had been collected in the container placed below the cube. Hence
it was concluded that most of the test substance sample had evaporated during the test.
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Page 15 of 27
September 2009
Test Facility
NICNAS
NOTOX (2009a)
Explosive Properties
Method
Remarks
Test Facility
EC Directive 92/69/EEC A.14 Explosive Properties.
The test substance is not explosive as it is not thermally sensitive, shock sensitive or
friction sensitive.
NOTOX (2009a)
Oxidizing Properties
Method
Remarks
Test Facility
Not explosive
No oxidizing properties
Expert Statement
The molecular structure of the test substance suggests that it is unlikely to have oxidising
properties.
NOTOX (2009a)
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Page 16 of 27
September 2009
NICNAS
APPENDIX B: TOXICOLOGICAL INVESTIGATIONS
B.1.
Acute toxicity – oral
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 423 Acute Oral Toxicity – Acute Toxic Class Method.
EC Directive 92/69/EEC B.1tris Acute Oral Toxicity – Acute Toxic Class
Method.
Rat/Wistar Crl:WI (Han) (outbred, SPF quality)
Propylene glycol
No significant protocol deviations.
Species/Strain
Vehicle
Remarks - Method
RESULTS
Dose
mg/kg bw
2000
2000
LD50
Signs of Toxicity
Number and Sex
of Animals
3F
3F
Mortality
0
0
>2000 mg/kg bw
Hunched posture, uncoordinated movements and/or piloerection were
noted in all animals on Day 1. Yellow faeces were noted in all animals on
Days 2 or 3.
None
Effects in Organs
CONCLUSION
The notified chemical is of low toxicity via the oral route.
TEST FACILITY
NOTOX (2008a)
B.2.
Acute toxicity – dermal
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 402 Acute Dermal Toxicity.
EC Directive 92/69/EEC B.3 Acute Toxicity (Dermal).
Rat/Wistar Crl:WI (Han) (outbred, SPF quality)
Propylene glycol
Occlusive
No significant protocol deviations.
Species/Strain
Vehicle
Type of dressing
Remarks - Method
RESULTS
Dose
mg/kg bw
2000
2000
LD50
Signs of Toxicity - Local
Signs of Toxicity - Systemic
Effects in Organs
Remarks - Results
Number and Sex
of Animals
5M
5F
Mortality
0
0
>2000 mg/kg bw
Scales and/or scabs were observed in the majority of the animals during
the observation period.
Chromodacryorrhoea was observed in two animals on Day 1. Given the
short duration of this symptom and the absence of any associated signs, it
was considered to be of no toxicological significance.
No toxicologically significant effects.
Yellow staining of the treated skin was noted in all animals during most
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Page 17 of 27
September 2009
NICNAS
of the observation period. One female animal also showed yellow staining
of other areas of the body on days 2 and 3. This was likely to be due to
the staining properties of the test substance.
CONCLUSION
The notified chemical is of low toxicity via the dermal route.
TEST FACILITY
NOTOX (2008b)
B.3.
Irritation – skin
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 404 Acute Dermal Irritation/Corrosion.
EC Directive 92/69/EEC B.4 Acute Toxicity (Skin Irritation).
Rabbit/New Zealand White
3 males
Water/ethanol (50% v/v)
72 hr
Semi-occlusive.
No significant protocol deviations.
Species/Strain
Number of Animals
Vehicle
Observation Period
Type of Dressing
Remarks - Method
RESULTS
Remarks - Results
No skin irritation effects were observed in the animals during the
observation period.
Yellow staining of the treated skin was observed in all animals throughout
the observation period. This did not interfere with the scoring of the skin
reactions.
CONCLUSION
The notified chemical is non-irritating to the skin.
TEST FACILITY
NOTOX (2008c)
B.4.
Irritation – eye
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 405 Acute Eye Irritation/Corrosion.
EC Directive 92/69/EEC B.5 Acute Toxicity (Eye Irritation).
Rabbit/New Zealand White
3 males
72 hr
No significant protocol deviations.
Species/Strain
Number of Animals
Observation Period
Remarks - Method
RESULTS
Lesion
Mean Score*
Animal No.
Maximum
Value
Maximum
Duration of Any
Effect
1
2
3
Conjunctiva: redness
0.3 1.0 0.3
2
<72hr
Conjunctiva: chemosis
0
0
0
1
<24hr
Conjunctiva: discharge
0.3
0
0
1
<48hr
Corneal opacity
0
0
0
0
Iridial inflammation
0
0
0
0
*Calculated on the basis of the scores at 24, 48, and 72 hours for EACH animal.
FULL PUBLIC REPORT: LTD/1414
Maximum Value at End
of Observation Period
0
0
0
0
0
Page 18 of 27
September 2009
Remarks - Results
NICNAS
Remnants of the test substance were present in the eye and/or on the
outside of the eyelids during the observation time. Yellow staining of fur
on the head and paws, caused by the test substance was also noted.
CONCLUSION
The notified chemical is slightly irritating to the eye.
TEST FACILITY
NOTOX (2008d)
B.5.
Skin sensitisation – mouse local lymph node assay (LLNA)
TEST SUBSTANCE
Notified chemical
METHOD
Species/Strain
Vehicle
Remarks - Method
OECD TG 429 Skin Sensitisation: Local Lymph Node Assay.
Mouse/CBA strain, inbred, SPF quality
Methyl ethyl ketone
No significant protocol deviations. In the preliminary irritation study the
50% material did not remain adhered to the ear, therefore 25% was
chosen as the highest concentration for the main study.
RESULTS
Concentration
(% w/w)
Test Substance
0 (vehicle control)
5
10
25
Positive Control
(-Hexylcinnamaldehyde)
5
10
25
Remarks - Results
Proliferative response
(DPM/lymph node)
Stimulation Index
(Test/Control Ratio)
369 ± 85
419 ± 72
523 ± 72
496 ± 101
1.0
1.1
1.4
1.3
445 ± 127
930 ± 141
2649 ± 505
1.0
2.0
5.7
Yellow staining by the test substance or its remnants prevented scoring for
erythema. No oedema was observed in any of the animals examined.
CONCLUSION
There was no evidence of induction of a lymphocyte proliferative
response indicative of skin sensitisation to the notified chemical.
TEST FACILITY
NOTOX (2008e)
B.6.
Repeat dose toxicity
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 407 Repeated Dose 28-day Oral Toxicity Study in Rodents.
EC Directive 96/54/EC B.7 Repeated Dose (28 Days) Toxicity (Oral).
Rat/Crl:CD(SD)
Oral – gavage
Total exposure days: 28 days
Dose regimen: 7 days per week
Post-exposure observation period: 14 days
Test substance was suspended in 5 %w/v gum arabic solution
No significant protocol deviations. Dose levels were chosen on the basis
of a preliminary 14 day repeated oral dose toxicity study.
Species/Strain
Route of Administration
Exposure Information
Vehicle
Remarks - Method
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Page 19 of 27
September 2009
NICNAS
RESULTS
Group
Number and Sex
of Animals
5M; 5F
5M; 5F
5M; 5F
5M; 5F
5M; 5F
5M; 5F
control
low dose
mid dose
high dose
control recovery
high dose recovery
Dose
mg/kg bw/day
0
50
250
1000
0
1000
Mortality
0/10
0/10
0/10
0/10
0/10
0/10
Mortality and Time to Death
No mortality occurred during the study.
Clinical Observations
Yellow stool was continuously observed in both sexes of the mid and high dose groups. In the high dose
recovery group yellow stool was only observed up to three days after termination of the dosing period in both
sexes. This was considered to be a treatment-related effect, though not of toxicological significance, as it
reflected the colour of the test substance.
Remarks – Results
No other toxicologically significant changes were observed in animals during the study.
CONCLUSION
The No Observed Adverse Effect Level (NOAEL) was established as 1000 mg/kg bw/day in this study, based
on the absence of toxicologically significant effects.
TEST FACILITY
CERI (2008a)
Genotoxicity – bacteria
B.7.
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 471 Bacterial Reverse Mutation Test.
EC Directive 2000/32/EC B.13/14 Mutagenicity – Reverse Mutation Test
using Bacteria.
Pre incubation procedure
S. typhimurium: TA1535, TA1537, TA98, TA100
E. coli: WP2uvrA
S9-mix (uninduced male Golden Syrian Hamster liver S9-mix in
modified co-factors)
a) With metabolic activation:
3 - 333 µg/plate
b) Without metabolic activation: 3 - 333 µg/plate
Dimethyl sulfoxide
The method incorporated the Prival and Mitchell modification for azo
compounds (Prival MJ and Mitchell VD 1982).
The concentrations chosen for the main study were based on the
observation of precipitation during the range finding study. The highest
concentration used in the main study was that at which limited solubility
was shown.
Species/Strain
Metabolic Activation System
Concentration Range in
Main Test
Vehicle
Remarks - Method
RESULTS
Metabolic
Activation
Test Substance Concentration (µg/plate) Resulting in:
Cytotoxicity in
Cytotoxicity in
Precipitation*
Genotoxic Effect
Preliminary Test
Main Test
Absent
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September 2009
NICNAS
Test 1
>5000
>333
>100
Negative
Test 2
>333
>100
Negative
Present
Test 1
>5000
>333
>100
Negative
Test 2
>333
>100
Negative
*Precipitation observed at the end of the incubation period. Note that precipitation was observed at the start of
the incubation period at concentrations of >10 µg/plate.
Remarks - Results
The test substance caused no visible reduction in the growth of the
bacterial background lawn or decrease in the number of revertants at any
dose level in all tester strains in the presence and absence of S9-mix. No
significant increases in the frequency of revertant colonies were recorded
for any of the bacterial strains, with any dose of the test material.
The Prival-Mitchell modification positive control, Congo Red, used in
the test induced marked increases in the frequency of the TA98 revertant
colony with metabolic activation only. Thus, the sensitivity of the assay
and the efficacy of the uninduced hamster liver S9-mix was validated.
CONCLUSION
The notified chemical was not mutagenic to bacteria under the conditions
of the test.
TEST FACILITY
NOTOX (2008f)
B.8.
Genotoxicity – in vitro
TEST SUBSTANCE
Notified chemical
METHOD
Species/Strain
Cell Type/Cell Line
Metabolic Activation System
OECD TG 473 In vitro Mammalian Chromosome Aberration Test.
Chinese hamster
Chinese hamster lung fibroblasts (CHL/IU cells)
Rat liver S9-mix induced by a combination of phenobarbital and 5,6benzoflavone.
0.5 w/v% carboxymethyl cellulose sodium salt (CMC) solution
During the 24 hour continuous treatment test the medium was exchanged
2 hours before the end of the culture using the same procedure as that of
the short term treatment (due to difficulties in analysis of the
chromosomes caused by precipitation of the test substance).
Vehicle
Remarks - Method
In each treatment, the highest dose for observation of chromosomal
aberration was selected at the highest dose that the analysis of the
chromosomal aberration was possible (at higher concentrations cells were
condensed, spread of the chromosomes was poor, and the test substance
was observed to precipitate on the chromosomes).
Metabolic
Test Substance Concentration (μg/mL)
Activation
Absent
Test 1
78.1, 156*, 313*, 625*, 1250, 2500, 5000
Test 2
Present
Test 1
78.1, 156*, 313*, 625*, 1250*, 2500, 5000
Test 2
19.5*, 39.1*, 78.1*, 156, 313, 625, 1250
*Cultures selected for metaphase analysis.
FULL PUBLIC REPORT: LTD/1414
Exposure
Period
Harvest
Time
6 hr
24 hr
6 hr
24 hr
24 hr
24 hr
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September 2009
NICNAS
RESULTS
Metabolic
Activation
Cytotoxicity in
Preliminary Test
Test Substance Concentration (µg/mL) Resulting in:
Cytotoxicity in
Precipitation*
Genotoxic Effect
Main Test
Absent
Test 1
1300
1800
All doses
Negative
Test 2
Present
Test 1
2100
>5000
All doses
Negative
Test 2
320
250
All doses
Negative
*Precipitation observed at the start and end of treatment. At the end of the culture, precipitation was observed at
≥156 µg/mL in both short term treatment tests.
Remarks - Results
The frequency of cells with structural or numerical aberrations was not
significantly increased following treatment with the test substance.
CONCLUSION
The notified chemical was not clastogenic to Chinese hamster lung
fibroblasts treated in vitro under the conditions of the test.
TEST FACILITY
CERI (2007)
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September 2009
NICNAS
APPENDIX C: ENVIRONMENTAL FATE AND ECOTOXICOLOGICAL INVESTIGATIONS
C.1.
Environmental Fate
C.1.1. Ready biodegradability
TEST SUBSTANCE
Notified chemical
METHOD
Inoculum
Exposure Period
Auxiliary Solvent
Analytical Monitoring
OECD TG 301 C Ready Biodegradability: Modified MITI Test (I).
Activated sludge
28 days
None
Oxygen consumption and HPLC
RESULTS
Day
7
28
Test substance
% Degradation
0
0
Aniline
Day
7
28
CONCLUSION
The notified chemical is not readily biodegradable.
TEST FACILITY
CERI (2008b)
% Degradation
52
76
C.1.2. Bioaccumulation
Remarks
C.2.
A bioaccumulation test was not conducted because of the lack of a
sufficiently sensitive analytical method, but the notifier argues that the
notified chemical is not expected to have potential for bioaccumulation
because bioconcentration factors for analogue pigments are < 10. While
supporting test reports were not presented, the notified chemical is not
expected to bioconcentrate in fish because of its very limited affinity for
the lipid phase of living organisms, as has been discussed in the screening
assessment of Pigment Red 187 (Environment Canada and Health
Canada, 2008).
Ecotoxicological Investigations
C.2.1. Acute toxicity to fish
TEST SUBSTANCE
Notified chemical
METHOD
Species
Exposure Period
Auxiliary Solvent
Water Hardness
Analytical Monitoring
Remarks – Method
OECD TG 203 Fish, Acute Toxicity Test – static..
Oryzias latipes (Medaka)
96 hours
None
28 mg CaCO3/L
HPLC of tetrahydrofuran extracts of filtered samples
The solubility of the test substance in the test medium was 0.014 mg/L
RESULTS
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Page 23 of 27
September 2009
NICNAS
Concentration mg/L
Nominal
Actual
0
0
100
0.024
LC50
NOEC (or LOEC)
Remarks – Results
Number of Fish
8
8
1h
0
0
24 h
0
0
Mortality
48 h 72 h
0
0
0
0
96 h
0
0
> 0.024 mg/L at 96 hours.
0.024 mg/L at 96 hours.
No sublethal effects were observed.
CONCLUSION
The notified chemical is not toxic to fish at concentrations up to the
solubility limit.
TEST FACILITY
CERI (2008c)
C.2.2. Acute toxicity to aquatic invertebrates
TEST SUBSTANCE
Notified chemical
METHOD
Species
Exposure Period
Auxiliary Solvent
Water Hardness
Analytical Monitoring
Remarks – Method
OECD TG 202 Daphnia sp. Acute Immobilisation Test - static.
Daphnia magna
48 hours
None
28 mg CaCO3/L
HPLC of tetrahydrofuran extracts of filtered samples
The solubility of the test substance in the test medium was 0.017 mg/L
RESULTS
Concentration mg/L
Nominal
Actual
0
0
100
0.031
LC50
NOEC (or LOEC)
Remarks - Results
Number of D. magna
20
20
Number Immobilised
24
48 h
0
0
0
0
> 0.031 mg/L at 48 hours
0.031 mg/L at 48 hours
No abnormal responses were observed.
CONCLUSION
The notified chemical is not toxic to daphnids at concentrations up to the
solubility limit.
TEST FACILITY
CERI (2008d)
C.2.3. Algal growth inhibition test
TEST SUBSTANCE
Notified chemical
METHOD
Species
Exposure Period
Concentration Range
OECD TG 201 Alga, Growth Inhibition Test..
Pseudokirchneriella subcapitata
72 hours
Nominal:
0 and 100 mg/L
Actual:
0 and 0.0043 mg/L
None
Soft water (OECD culture medium)
HPLC of tetrahydrofuran extracts of filtered samples.
The solubility of the test substance in the test medium was 0.029 mg/L
Auxiliary Solvent
Water Hardness
Analytical Monitoring
Remarks - Method
RESULTS
FULL PUBLIC REPORT: LTD/1414
Page 24 of 27
September 2009
NICNAS
Biomass
Growth
NOEC
mg/L
0.0043
EbC50
mg/L at 72 h
> 0.0043
Remarks - Results
ErC50
mg/L at 72 h
>0.0043
NOEC
mg/L
0.0043
Cell growth in controls met the validity criterion.
CONCLUSION
The notified chemical is not toxic to freshwater green algae at
concentrations up to the solubility limit.
TEST FACILITY
CERI (2008e)
C.2.4. Inhibition of microbial activity
TEST SUBSTANCE
Notified chemical
METHOD
OECD TG 209 Activated Sludge, Respiration Inhibition Test.
EC Directive 88/302/EEC C.11 Biodegradation: Activated Sludge
Respiration Inhibition Test
Activated sludge
3 hours
Nominal:
100 mg/L
Sonication was used to facilitatate dispersion of the poorly soluble test
substance in the test medium. Contact was optimised by stirring and
continuous aeration.
Inoculum
Exposure Period
Concentration Range
Remarks – Method
RESULTS
IC50
NOEC
Remarks – Results
> 100 mg/L
100 mg/L
CONCLUSION
The test substance does not inhibit the respiration of activated sludge.
TEST FACILITY
NOTOX (2008g)
FULL PUBLIC REPORT: LTD/1414
Page 25 of 27
September 2009
NICNAS
BIBLIOGRAPHY
Brown MA and DeVito SC (1993). Predicting azo dye toxicity, Critical Reviews in Environmental Science and
Technology, 23(3): 249-324.
Chilworth (2008) Particle size analysis on a sample of Y-513. Project no 486797, July 2008, Chilworth
Technology Limited, Southampton, UK (unpublished report provided by notifier).
DFG (1988) Deutsche Forschungsgemeinschaft. List of MAK and BAT Values 1988. VCH Publishers,
Weinheim.
Environment Canada and Health Canada (2008) Screening Assessment for the Challenge: 4-[[5-[[[4(Aminocarbonyl)phenyl]amino]carbonyl]-2-methoxyphenyl]azo]-N-(5-chloro-2,4-dimethoxyphenyl)-3hydroxynaphthalene-2-carboxamide
(Pigment
Red
187)
July
2008
(http://www.ec.gc.ca/substances/ese/eng/challenge/batch1/batch1_59487-23-9_en.pdf
accessed 22 May
2009).
European Commission (2003) Technical Guidance Document on Risk Assessment in Support of Commission
Directive 93/67/EEC on Risk Assessment for New Notified Substances and Commission Regulation (EC)
No 1488/94 on Risk Assessment for Existing Substances and Directive 98/8/EC of the European Parliament
and of the Council Concerning the Placing of Biocidal Products on the Market – Part I. Institute for Health
and Consumer protection, European Chemicals Bureau, European Communities.
CERI (2007) Chromosomal Aberration Test of Y-513 Using Cultured Mammalian Cells. Study no K06-1206,
August 2007. Hita Laboratory, Chemicals Evaluation and Research Institute, Japan. (Unpublished report
provided by notifier).
CERI (2008a) Twenty-Eight Day Repeated-Dose Oral Toxicity Study of Y-513 in Rats. Study no B11-0846,
March 2008. Hita Laboratory, Chemicals Evaluation and Research Institute, Japan. (Unpublished report
provided by notifier).
CERI (2008b) Biodegradation Study of Y-513 by Microorganisms. Study no 14914, report dated 27 June 2008.
Kurume Laboratory, Chemicals Evaluation and Research Institute, Japan. (Unpublished report provided by
notifier).
CERI (2008c) A 96-hour Acute Toxicity Study of Y-513 with Medaka. Study no 94349, report dated 8 October
2008. Kurume Laboratory, Chemicals Evaluation and Research Institute, Japan. (Unpublished report
provided by notifier).
CERI (2008d) A 48-hour Acute Immobilisation Study of Y-513 with Daphnia magna. Study no 94348, report
dated 7 October 2008. Kurume Laboratory, Chemicals Evaluation and Research Institute, Japan.
(Unpublished report provided by notifier).
CERI (2008e) Algae Growth Inhibition Study of Y-513 with Pseudokirchneriella subcapitata. Study no 948078,
report dated 9 December 2008. Kurume Laboratory, Chemicals Evaluation and Research Institute, Japan.
(Unpublished report provided by notifier).
IARC (1987) International Agency for Research on Cancer. Benzidine-Based Dyes (Group 2A). Supplement 7,
pg 125
NOHSC (1994) National Code of Practice for the Labelling of Workplace Substances [NOHSC:2012(1994)].
National Occupational Health and Safety Commission, Canberra, Australian Government Publishing
Service.
NOHSC (2003) National Code of Practice for the Preparation of Material Safety Data Sheets, 2nd edition
[NOHSC:2011(2003)]. National Occupational Health and Safety Commission, Canberra, Australian
Government Publishing Service.
NOHSC (2004) Approved Criteria for Classifying Hazardous Substances, 3rd edition [NOHSC:1008(2004)].
National Occupational Health and Safety Commission, Canberra, AusInfo.
NOTOX (2008a) Assessment of Acute Oral Toxicity with Y-513 in the Rat (Acute Toxic Class Method). Project
no 486805, November 2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands. (Unpublished report provided
by notifier).
NOTOX (2008b) Assessment of Acute Dermal Toxicity with Y-513 in the Rat. Project no 486806, December
2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands. (Unpublished report provided by notifier).
FULL PUBLIC REPORT: LTD/1414
Page 26 of 27
September 2009
NICNAS
NOTOX (2008c) Primary Skin Irritation/Corrosion Study with Y-513 in the Rabbit (4-hour semi-occlusive
application). Project no 486807, November 2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands.
(Unpublished report provided by notifier).
NOTOX (2008d) Acute Eye Irritation/Corrosion Study with Y-513 in the Rabbit. Project no 486808, November
2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands. (Unpublished report provided by notifier).
NOTOX (2008e) Assessment of Contact Hypersensitivity to Y-513 in the Mouse (Local Lymph Node Assay).
Project no 486809, November 2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands. (Unpublished report
provided by notifier).
NOTOX (2008f) Evaluation of the Mutagenic Activity of Y-513 in the Salmonella Typhimurium Reverse
Mutation Assay and the Escherichia Coli Reverse Mutation Assay (with independent repeat). Project no
486811, January 2008, NOTOX BV, ’s-Hertogenbosch, the Netherlands. (Unpublished report provided by
notifier).
NOTOX (2008g) Activated Sludge Respiration Inhibition test with Y-513. Project no 486812, May 2008.
NOTOX BV, the Netherlands. (Unpublished report provided by notifier).
NOTOX (2009a) Determination of Physico-Chemical Properties of Y-513. Project no 486786, report dated 23
March 2009. NOTOX BV, the Netherlands. (Unpublished report provided by notifier).
NOTOX (2009b) Toxicokinetic assessment of Y-513. Project no 486810, March 2009, NOTOX BV, the
Netherlands. (Unpublished report provided by notifier).
Øllgaard H, Frost L, Galster J and Hansen OC (1998). Survey of azo-colorants in Denmark: Consumption, use,
health and environmental aspects. Danish Technological Institute, Environment, Danish Environmental
Protection Agency.
Prival MJ and Mitchell VD (1982) Analysis of a method for testing azo dyes for mutagenic activity in
Salmonella typhimurium in the presence of flavin mononucleotide and hamster liver S9. Mutat Res. 97(2):
103-16.
SCCNFP (2002) The Safety Review Of The Use Of Certain Azo-Dyes In Cosmetic Products: Opinion Of The
Scientific Committee On Cosmetic Products And Non-Food Products Intended For Consumers.
SCCNFP/0495/01 (prepared in the context of Directive 76/768/EEC).
US EPA (2002). TSCA New Chemicals Program (NCP) Chemical Categories. 1200 Pennsylvania Avenue,
N.W, Washington, D.C. 20460 http://www.epa.gov/opptintr/newchems/pubs/cat02.htm.
FULL PUBLIC REPORT: LTD/1414
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